Compositions and combinations of organophosphorus bioscavengers and hyaluronan-degrading enzymes, and methods of use

Abstract

Provided are compositions and combinations containing an organophosphorus bioscavenger and a hyaluronan-degrading enzyme. The provided compositions and combinations can be used to treat or prevent organophosphorus poisoning, including nerve agent poisoning and pesticide poisoning.

1 . A method for treating or preventing organophosphorus poisoning by administering a composition, comprising an organophosphorus (OP) bioscavenger and a hyaluronan-degrading enzyme. 
     
     
         2 . The method of  claim 1 , wherein the composition is administered in a single dose. 
     
     
         3 . The method of  claim 1 , wherein the composition is administered in multiple doses. 
     
     
         4 . The method of  claim 1 , wherein the organophosphorus bioscavenger is an esterase, cholinesterase, paraoxonase (PON), aryldialkylphosphatase or diisopropylfluorophosphatase (DFPase). 
     
     
         5 . The method of  claim 1 , wherein the organophosphorus bioscavenger is selected from among acetylcholinesterase (AChE), butyrylcholinesterase (BChE), prolidase, organophosphate acid anhydrolase (OPAA), phosphotriesterase, aryldialkylphosphatase, organophosphorus hydrolase (OPH), parathion hydrolase, diisopropylfluorophosphatase (DFPase), organophosphorus acid anhydrase, sarinase and paraoxonase (PON) and an active portion thereof or a variant thereof that exhibits at least 80% OP binding or inactivating activity. 
     
     
         6 . The method of  claim 1 , wherein the organophosphorus bioscavenger has the sequence of amino acids set forth in any of SEQ ID NOS:214-256 and 258-301, an active portion thereof or a variant thereof that has at least 80% sequence identity to any of SEQ ID NOS:214-256 and 258-301. 
     
     
         7 . The method of  claim 1 , wherein the organophosphorus bioscavenger is butyrylcholinesterase that has the sequence of amino acids set forth in SEQ ID NO:236, or is an active portion thereof or is a variant thereof that has at least 85% sequence identity to the sequence of amino acids set forth in SEQ ID NO:236. 
     
     
         8 . The method of  claim 1 , wherein the organophosphorus bioscavenger is modified with a polymer. 
     
     
         9 . The method of  claim 8 , wherein the polymer is a polyethylene glycol (PEG). 
     
     
         10 . The method of  claim 1 , wherein the organophosphorus bioscavenger is linked directly or indirectly via a linker to an immunoglobulin, immunoglobulin domain, albumin, transferrin, or transferrin receptor protein. 
     
     
         11 . The method of  claim 1 , wherein the hyaluronan-degrading enzyme is a hyaluronidase or a chondroitinase, or a variant or a truncated form thereof that exhibits hyaluronan-degrading activity. 
     
     
         12 . The method of  claim 1 , wherein the hyaluronan-degrading enzyme is a hyaluronidase that is a PH20 or a variant or a truncated form thereof that exhibits hyaluronidase activity. 
     
     
         13 . The method of  claim 1 , wherein the hyaluronan-degrading enzyme is a truncated human PH20 that consists of a sequence of amino acids set forth in SEQ ID NO:1 that contains a C-terminal truncation after amino acid position 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499 or 500 of the sequence of amino acids set forth in SEQ ID NO:1, or is a variant thereof that exhibits at least 85% sequence identity to a sequence of amino acids that contains a C-terminal truncation after amino acid position 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499 or 500 of the sequence of amino acids set forth in SEQ ID NO:1 and exhibits hyaluronidase activity. 
     
     
         14 . The method of  claim 13 , wherein the hyaluronan-degrading enzyme has a sequence of amino acids that contains at least amino acids 36-464 of SEQ ID NO:1, or has a sequence of amino acids that has at least 85% sequence identity to a sequence of amino acids that contains at least amino acids 36-464 of SEQ ID NO:1 and exhibits hyaluronidase activity at neutral pH. 
     
     
         15 . The method of  claim 1 , wherein the hyaluronan-degrading enzyme comprises the sequence of amino acids set forth in any of SEQ ID NOS:4-9 or a sequence of amino acids that exhibits at least 85% sequence identity to any of SEQ ID NOS:4-9. 
     
     
         16 . The method of  claim 1 , wherein the hyaluronan-degrading enzyme is modified with a polymer. 
     
     
         17 . The method of  claim 16 , wherein the polymer is a polyethylene glycol (PEG). 
     
     
         18 . The method of  claim 1 , wherein the organophosphorus bioscavenger is present in the composition at a concentration of between or between about 1 to 1000 μg/mL, 0.5 to 50 mg/mL, 1 to 1000 mg/mL, 50 to 1500 mg/mL, or 100 to 750 mg/mL. 
     
     
         19 . The method of  claim 1 , wherein the hyaluronan-degrading enzyme is present in the composition at a concentration between or between about 10 U/mL to 100,000 U/mL, 1000 U/mL to 50,000 U/mL, 5,000 U/mL to 20,000 U/mL or 10 U/mL to 10,000 U/mL. 
     
     
         20 . The method of  claim 1 , wherein the hyaluronan-degrading enzyme is present in the composition at a concentration between or about between 10 U/mL to 5000 U/mL. 
     
     
         21 . The method of  claim 1 , wherein the organophosphorus bioscavenger and hyaluronan-degrading enzyme are administered by subcutaneous administration, intramuscular administration, intralesional administration or intradermal administration. 
     
     
         22 . The method of  claim 1 , wherein the organophosphorus bioscavenger and hyaluronan-degrading enzyme are administered by intramuscular administration. 
     
     
         23 . The method of  claim 1 , wherein the composition is administered between at or about 6 to 48 hours, 6 to 36 hours, 6 to 24 hours, 12 to 48 hours, 12 to 36 hours, 12 to 24 hours, 24 to 48 hours, 24 to 36 hours, or at least or at least about 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 24, 30, 36, 42 or 48 hours before exposure to the organophosphorus compound. 
     
     
         24 . The method of  claim 1 , further comprising administering another pharmaceutical agent selected from among carbamates, anti-muscarinics, cholinesterase reactivators and anti-convulsives. 
     
     
         25 . A method for treating or preventing organophosphorus poisoning by administering a combination, comprising:
 administering a first composition containing an organophosphorus bioscavenger; and   administering a second composition containing a hyaluronan-degrading enzyme.   
     
     
         26 . The method of  claim 25 , wherein the organophosphorus bioscavenger and hyaluronan-degrading enzyme are administered by subcutaneous administration, intramuscular administration, intralesional administration or intradermal administration. 
     
     
         27 . The method of  claim 25 , wherein the organophosphorus bioscavenger and hyaluronan-degrading enzyme are administered by intramuscular administration. 
     
     
         28 . The method of  claim 25 , wherein the combination is administered between at or about 6 to 48 hours, 6 to 36 hours, 6 to 24 hours, 12 to 48 hours, 12 to 36 hours, 12 to 24 hours, 24 to 48 hours, 24 to 36 hours, or at least or at least about 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 24, 30, 36, 42 or 48 hours before exposure to the organophosphorus compound. 
     
     
         29 . The method of  claim 25 , wherein the hyaluronan-degrading enzyme is a hyaluronidase that is a PH20 or a variant or a truncated form thereof that exhibits hyaluronidase activity.